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The N-glycan on Asn54 affects the atypical N-glycan composition of plant-produced interleukin-22, but does not influence its activity
- Source :
- Plant Biotechnology Journal, 14(2), 670-681, Wilbers, R H P, Westerhof, L B, Reuter, L J, Castilho, A, van Raaij, D R, Nguyen, D L, Lozano-Torres, J L, Smant, G, Hokke, C H, Bakker, J & Schots, A 2016, ' The N-glycan on Asn54 affects the atypical N-glycan composition of plant-produced interleukin-22, but does not influence its activity ', Plant Biotechnology Journal, vol. 14, no. 2, pp. 670-681 . https://doi.org/10.1111/pbi.12414, Plant Biotechnology Journal 14 (2016) 2
- Publication Year :
- 2016
- Publisher :
- Wiley, 2016.
-
Abstract
- Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. IL-22 is an unusual cytokine as it does not act directly on immune cells. Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. Expression levels of IL-22 were up to 5.4 μg/mg TSP, and N-glycan analysis revealed the presence of the atypical Lewis A structure. Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. Also, core α1,6-fucoylation did not improve the biological activity of IL-22. The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22.
- Subjects :
- 0301 basic medicine
PNGase F
Nicotiana
Glycan
Glycosylation
Core α1,6-fucose
Nicotiana benthamiana
Plant Science
Core α1
N-glycosylation
Interleukin-22
Glyco-engineering
Interleukin 22
03 medical and health sciences
N-linked glycosylation
Polysaccharides
Tobacco
Animals
Humans
6-fucose
Laboratorium voor Nematologie
chemistry.chemical_classification
core 1
biology
Lewis A
EPS-2
Interleukins
Biological activity
biology.organism_classification
Plants, Genetically Modified
Plant Leaves
carbohydrates (lipids)
030104 developmental biology
Drosophila melanogaster
HEK293 Cells
Biochemistry
chemistry
Metabolic Engineering
biology.protein
Protein folding
Laboratory of Nematology
Asparagine
Glycoprotein
Agronomy and Crop Science
Biotechnology
Subjects
Details
- Language :
- English
- ISSN :
- 14677652 and 14677644
- Volume :
- 14
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Plant Biotechnology Journal
- Accession number :
- edsair.doi.dedup.....5537f68c2587cf304d52cd77ebd5b911